Layered Double Hydroxide Nanoparticles with Osteogenic Effects as miRNA Carriers to Synergistically Promote Osteogenesis of MSCs.

Inefficient differentiation and poor engraftment hinder the clinical applications of mesenchymal stem cell (MSC)-based cell therapies in regenerative medicine. Layered double hydroxide (LDH) nanoparticles are sheet-like materials with desirable biocompatibility and anion-exchange properties and have been widely applied as drug and nucleotide carriers in the field of tissue repair. However, few studies have focused on the biological effects of LDH itself. In this study, we demonstrated the novel function of LDH in stimulating osteogenic differentiation of bone marrow-derived MSCs (BMSCs). The expression of osteogenic-related genes, alkaline phosphatase (ALP) activity, and calcium deposits were significantly increased after LDH treatment. Mechanistic analysis performed with RNA sequencing revealed that LDH promoted osteogenesis by targeting the LGR5/β-catenin axis. LDH also inactivated IKK/NF-κB signaling under LPS-triggered inflamed conditions, suggesting the dual benefits of LDH in enhancing bone regeneration and alleviating the inflammatory response. Furthermore, we utilized LDH as the transport vehicle of the osteoinductive miRNA let-7d to synergistically regulate BMSCs toward the osteoblastic lineage. The LDH/let-7d complex resulted in a better induction of osteogenesis than LDH alone. For cell transplantation, BMSCs were seeded in LDH/let-7d-incorporated fibrin scaffolds, which proved enhanced osteoinduction capability in the subcutaneous ectopic osteogenesis model in nude mice. Taken together, this study provides a novel strategy for effective and synergistic improvement of osteogenesis via LDH-mediated delivery of miRNA let-7d, thus shedding light on the future application of LDH in regenerative medicine.